Preparation method of alginate microparticles capable of inducing tumor immune response

A technology of alginate and immune response, applied in the direction of anti-tumor drugs, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the difficult standardization of preparation methods, different curative effects, and affect the application of DC vaccines, etc. question

Inactive Publication Date: 2011-07-13
HAINAN MEDICAL COLLEGE
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method of preparing vaccines is relatively complicated and requires certain skills and experience. The preparation method is difficult to standardize, the cost of treatment is also high, and the efficacy often varies with different laboratories.
This "workshop" preparation method has seriously affected the further clinical application of DC vaccines

Method used

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  • Preparation method of alginate microparticles capable of inducing tumor immune response
  • Preparation method of alginate microparticles capable of inducing tumor immune response
  • Preparation method of alginate microparticles capable of inducing tumor immune response

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Embodiment Construction

[0021] The present invention will be further described below in conjunction with preferred embodiments, but the present invention is by no means limited to the following embodiments.

[0022] (1) Construction and identification of eukaryotic expression plasmid of MIP-3α

[0023] We purchased the cloning plasmid pORF5-mMIP3α from InvioGen, and designed a pair of PCR primers (upstream primer: 5'-CGT CTA GAT TAC ATC TTC TTG ACT CTT AGG C-3'; Downstream primer: 5'-AAGAAT TCATGG CCT GCG GTG GAAA-3'), using the plasmid pORF5-mMIP3α as a template, using PCR method (94℃ 30sec; 56℃ 1min; 72℃ 3.5min; total 30 Cycle) amplify the mouse MIP-3α gene ( figure 1 B). After that, using genetic engineering technology, the MIP-3α gene amplified by PCR was ligated into the eukaryotic expression plasmid pcDNA3.1(+) through restriction digestion and ligation, and restriction digestion ( figure 1 C) and gene sequencing ( figure 1 D) Method for analysis to obtain the correct recombinant eukaryotic express...

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Abstract

The invention provides a preparation method of alginate microparticles capable of inducing a tumor immune response, which comprises the steps of: establishing an MIP-3alpha (Macrophage Inflammatory Protein-3alpha) eukaryon expression plasmid and transforming tumor cells by using a gene engineering technology, and sieving to obtain the tumor cells stably expressing the MIP-3alpha; meanwhile, culturing escherichia coli and dissolving the escherichia coli in an alginate solution, disinfecting by means of high temperature and high pressure, uniformly mixing the MIP-3alpha tumor cells in the escherichia coli-alginate solution, atomizing the MIP-3alpha tumor cell-escherichia coli-alginate solution by using an atomizer, spraying atomized particles into a calcium chloride solution, and filtering by using a gauze element to obtain injectable MIP-3alpha tumor cell-escherichia coli-alginate microparticles, which can express secretion MIP-3alpha protein in an in-vitro culture environment. After being injected into a mouse mantle cavity, the microparticles are demonstrated to be effective in inhibiting tumor growth and transfer in a tumor model and have obvious effects on treating tumors.

Description

Technical field [0001] The invention relates to an alginate microparticle, in particular to a preparation method of the alginate microparticle which can induce tumor immune response. Background technique [0002] The treatment of malignant tumors is still a problem in the world, and immunobiological therapy is the current research hotspot. Dendritic cells (DC) are the strongest known at present, and are the only full-time antigen presenting cell (APC) that can activate the native T cell (native T cell), with powerful activation CD8 + T-cytotoxic lymphocytes (CTL) and CD4 + The ability of T helper cells controls the process of immune response in the body and is the central link of tumor immune response. DCs are mainly distributed in bone marrow, peripheral blood, and organs and lymphoid tissues where antigens are exposed. DCs that have not been exposed to antigens are called immature DCs. Immature DCs have strong capture and processing. Antigen ability; when the DC captures and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/00A61K47/36A61K9/14A61P35/00A61P35/04
Inventor 谭光宏黄风迎黎岳南郭峻丽黄用豪王华
Owner HAINAN MEDICAL COLLEGE
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